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Int.J.Curr.Microbiol.App.Sci
(201
4) 3(9) 1183-
120
0
1183
Original Research Article
Distribution of Microorganisms in Water, Soils and Sediment
f
rom
Abattoir Wastes in Southern Nigeria
David N Ogbonna
*
Department of Applied and Environmental Biology Rivers State University
of Science and
Technology,
PMB
5080, Port Harcourt, Nigeria
*Corresponding author
A B S T R A C T
Introduction
The continuous drive to increase meat
production for the protein need of the ever
increasing world population has been
accompanied by some pollution problems
(Adesemoye
et al., 2006; Nafarnda et al
.,
2012). In Nigeria, the abattoir industry is an
important component of the livestock
indus
try providing domestic meat supply to
over 150 million people and employment
opportunities for teaming population
(Nafarnda
et al., 2012). Adeyemi and
Adeyemo (2007) reported that cities face
ISSN: 2319
-7706
Volume
3
Number
9
(201
4
) pp.
1183-
120
0
http://
www.ijcmas.com
Keyw ords
Abattoir
wastes,
Micro
-
organisms,
soil,
wastewater,
sediment,
public health
This study was carried out to determine the distribution of microorganisms
particularly those of public health importance in areas where abattoir activities are
in operation. Abattoirs located at Egbu in Imo State and Trans-Amadi in Port
Harcour
t, Rivers State was selected for sampling using standard analytical
methods. A total of twenty sampling points were established using Global
Positioning System (GPS). Sampling was carried out both in wet and dry seasons.
Result showed that soil samples from Trans-Amadi abattoir had highest levels of
total coliform and total
Vibrio
counts of 2.9x107cfu/g and 6.0x106 cfu/g,
respectively while surface water had the highest
Salmonella
and
Shigella
counts of
1.5x10
7 cfu/ml. In Egbu abattoir, statistical analysis using ANOVA revealed a
significant difference at 0.05 level in the total heterotrophic bacterial count from
sediment and those from soil, waste water and surface water samples. The
predominant bacterial genera identified were Pseudomonas, Bacillus,
Staphy
lococcus, Micrococcus, Lactobacillus, Streptococcus, Klebsiella, Vibrio,
Salmonella, Escherichia coli, Citrobacter, Acinetobacter, Serratia, Proteus,
Enterobacter, Shigella, Flavobacterium, Achromobacter
species. Pollution of water
resources by abattoir wa
stes might lead to destruction of primary producers and this
in turn leads to diminishing consumer populations in water. The consequences of
such
anthropogenic pollution can also lead to the transmission of diseases by
water
borne pathogens. Therefore, continuous monitoring should be maintained in
order to promote and maintain a safe working environment and ensure detection
when abnormalities that c
ould endanger both workers and environment occur.
Int.J.Curr.Microbiol.App.Sci
(201
4) 3(9) 1183-
120
0
1184
serious problems of high volume of wastes
from abattoir due to inadequate disposal
technologies and high cost of management.
In Nigeria, adequate abattoir waste
management is lacking in all public abattoirs
such that large solid wastes and untreated
effluents are common sites (Odeyemi, 1991;
Adeyemo, 2002; Adebowale
et
al., 2010)
unlike in developed countries where these
facilities are adequately provided
(Ogbonnaya, 2008). These abattoir wastes
could be a source of embarrassment since
conventional methods of waste management
have been grossly neglected (Adedipe, 2002;
Adeyemi and Adeyemo, 2007).
Wastewater or effluent generated from the
abattoir is characterized by the presence of a
high concentration of whole blood of
slaughtered food animals and suspended
particles of semi- digested and undigested
feeds within the stomach and intestine of
slaughtered and dressed food animals
(Coker
et al., 2001). In addition, there may
also be the presence of pathogenic
microorganisms, such as
Salmonella,
Escherichia
coli
(including serotype
0157:H7),
Shigella, parasite eggs and
amo
ebic cysts (Bull and Rogers, 2001;
Adebowale
et al., 2010) which are of public
health importance.
Also, several pathogenic bacteria and fungi
species has been isolated from abattoir
wastewater and surface water; including
Staphylococcus, Escherichia coli,
Streptococcus, Salmonella, Aspergillus,
Mucor, Saccharomyces
and
Penicillium
species (Coker et al., 2001; Adesomoye et
al
., 2006; Adebowale et al., 2010). These
pathogens might threaten public health by
migrating into ground water or surface
water; wind or vectors like animals, birds
and arthropods can transmit diseases from
these microorganisms (Mason, 1991;
Meadows, 1995; Gauri, 2004 Raheem and
Morenikeji, 2008).
Bacteria from abattoir waste discharged into
water columns can subsequently be
absorbed to sediments, and when the bottom
stream is disturbed, the sediment releases
the bacteria back into the water columns
presenting long term health hazards (Sherer
et al., 1992; Nafarnda et al., 2012).
Pathogens present in animal carcasses or
shed in animal wastes may include
rotaviruses, hepatatitis E virus, Salmonella
spp.,
E.coli
0157:H7,
Yersinia
enterocolitica,
Campylobacter
spp.,
Cryptosporidium parvum, and
Giardia
lamblia
(Sobsey et al., 2002). Fecal wastes
from domestic livestock in the abattoir are
excr
eted on the floors of the animal pens,
the accumulation of these fecal materials act
as a collection basin for pathogenic
microorganisms which may spread between
animals and man leading to zoonoses
(Adeyemo
et al., 2002). These zoonotic
pathogens can exceed millions to billions
per gram of feces, and may infect humans
through various routes such as contaminated
air, contact with livestock animals or their
waste products, swimming in water
impacted by animal feces, exposure to
potential vectors (such as flies, mosquitoes,
water fowl, and rodents), or consumption of
food or water contaminated by animal
wastes (Armand-
Lefevre
et al., 1998;
Schlech
et al
., 2005).
In Rivers state, wastes generated from a
slaughterhouse in Trans-Amadi abattoir,
Port Harcourt, Nigeria are channeled
directly into one of the tributaries of the
River Niger. This act could introduce enteric
pathogens e.g.
Bacillus
sp
.,
Escherichia
sp.,
etc and excess nutrients into the river,
resulting to eutrophication (Odeyemi, 1991;
Adeyemo
et al., 2002). These consequences
of anthropogenic pollution during abattoir
operations can lead to the transmission of
diseases by water borne pathogens,
eutrophication of water bodies,
Int.J.Curr.Microbiol.App.Sci
(201
4) 3(9) 1183-
120
0
1185
accumulation of toxic or recalcitrant
chemicals in the soil, destabilization of
ecological balance and negative effects on
human health (Amisu et al., 2003; Nafarnda
et al., 2012). Abu-
Ashour
et al (1994)
revealed that some bacteria also possess the
ability to attach to solid/substrate surfaces
by electrostatic hydrogen bonding and
hydrophobic interactions. After attachment,
they secrete slimy materials that can attract
other organisms and nutrients to the
interface. Attachment to surfaces benefits
microorganisms in several ways both on
nutritional and survival basis which
invariab
ly enhances bioremediation (Abu-
Ashour
et al., 1994). This study therefore is
aimed at assessing the distribution of
microorganisms in the various location sites
where abattoir operations are carried out,
considering the environmental and public
health imp
lications.
Materials and Methods
Study Area
The study was carried out in abattoirs
located at Egbu in Imo State; and Trans-
Amadi in Port Harcourt, Rivers State. Egbu
lies within longitude 05° 28.432 - 05°
29.802 N and latitude 007° 03.200 - 007°
04.215 E (Fig 1). This area Egbu has a
tropical climate. The average relative
humidity is about 80%. The inhabitants of
the areas are mainly farmers, civil servants,
petty traders and casual workers.
Port Harcourt is located on longitude 4°
48.442 - 4° 49.444 N and latitude 007°
02.303
007° 03.545E. The climate of Port
Harcourt falls within the sub equatorial
climate belt. Temperature and humidity are
high throughout the year. The area is ma
rked
by two distinct seasons, the wet and dry
seasons, with 70% of the annual rain fall
between April and August, while 22% is
spread in the three months of September to
November. However, the driest months are
from December to March. The river located
at
the abattoir in Egbu in Imo state is
popularly called the Otamiri River while the
river in Trans Amadi abattoir in Port
Harcourt is called the Oginigba creek.
Sampling Points
A total of twenty (20) sampling points were
considered for the study. The sam
pling
stations, sampling points codes, sampling
points coordinates and types of samples
collected are presented in Table 1. During
sample collection, Global Positioning
System (GPS) machine (Model GPS 76) was
used for the location of the sampling points.
Collection of Samples
Soil Samples
Soil samples were collected from four
different sampling points coded A, B, C and
D from a depth of 0-15cm using soil auger.
About 500g of bulked composite soil
samples was collected from points A, B and
C; then prepared using the method of
Ekundayo and Obuekwe (1997). Soil sample
from point D, which is about 400m from
Egbu and Trans Amadi abattoirs served as
control sample. The soil samples were
collected into labeled polyethylene bags and
transported to the laboratory in a cooler
packed with ice blocks for analysis.
Surface Water samples
Surface water samples were collected using
the method of Odokuma and Okpokwasili
(1993). The collection was carried out using
4 litre plastic bottles previously sterilized
with 70% alcohol 24 hours before the final
collection. The bottles were rinsed 3 to 4
times with the water sample before the final
Int.J.Curr.Microbiol.App.Sci
(201
4) 3(9) 1183-
120
0
1186
collection. The water samples were collected
along the course of the river at two different
points coded A and B. Point A is the
immed
iate point of discharge of the abattoir
wastes into the river, Point B is about 400m
upstream from Point A. The sample from
point A served as the test sample while that
from point B served as the control sample.
To collect the water sample, base of the
ste
rilized sample bottle was held with one
hand, the bottle was plunged about 30cm
below the water surface with the mouth of
the sample bottles positioned in an opposite
direction to water flow. The bottle was filled
with water sample leaving a gap of about
2cm and covered immediately as described
by Onyeagba and Umeham (2004).
Immediately after collection, the samples
were labeled and transported to the
laboratory in a cooler packed with ice blocks
for analysis.
Sediment samples
Sediment samples were collected from the
same sampling points where surface water
samples were collected using a grab
sampler. The sediment sample was scooped
from the grab s cup and transferred into
sterile sample bottle. The sample was
labeled and then transported to the
laboratory
in a cooler packed with ice blocks
for analysis.
Waste water Samples
Waste water samples were collected using
the method of Adesemoye et al. (2006).
Sterile 2.0 litre sample bottles were used to
aseptically draw part of the abattoir waste
water. The samples were collected at four
different points coded A, B, C and D as the
waste water was running off the drainage
system. About 500ml of the sample
collected from each point were pooled
together to get a composite sample. Control
samples were collected from water stored in
buckets used for washing meat and utensils
in the abattoirs. The samples were placed in
a cooler containing ice blocks and
transported immediately to the laboratory
for analysis.
Preparation of Samples
Sediment and Soil samples were processed
using the method of Adesemoye et al
.
(2006). Ten grams of the soil sample was
weighed and added to 90ml of sterile
distilled water to get an aliquot, similarly,
ten grams of the sediment sample was added
to 90ml of sterile distilled water to get an
aliquot. One milliliter of the aliquots, waste
water and surface water samples were then
serially diluted using the ten-fold serial
dilution method as described by Prescott
et
al
(2005).
Microbiological Analysis
The presence of various microorganisms in
the water samples from Otamiri River in
Imo state and Oginigba creek from Trans
Amadi abattoir in Port Harcourt were
identified using standard procedures. One
milliliter each of the waste water samples
was separately added to 9 ml of 0.1%
peptone water diluents to give a 10-3
dilution. After thorough shaking further
serial 10- fold (v/v) dilutions were made by
transferring 1 ml of the original solution to
freshly prepared peptone water diluents to a
range of 10-3 dilutions. Aliquots (0.1 ml) of
various dilutions were transferred to plates
of surface dried Nutrient agar in duplicate
and inoculated by spreading with flamed
glass spreaders and incubated at 370C for 24
hours. Aerobic bacteria were subjected to
further identification according to
determinative schemes of Cowan and Steel
(1994).
Int.J.Curr.Microbiol.App.Sci
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4) 3(9) 1183-
120
0
1187
Total heterotrophic bacterial counts: This
was determined with the nutrient agar using
the spread plate technique as described by
Prescott
et al (2005). Here 0.1ml of the
serially diluted samples was each inoculated
onto different sterile nutrient agar plates in
triplicates. The plates were incubated for 24
hours at 37oC. After incubation, colonies
that appeared on the plates were counted and
the mean expressed as cfu/ml for surface
water, wastewater and cfu/g for soil and
sediment samples.
Total coliform counts: The method of
Prescott
et al. (2005) was adopted where 0.1
milliliter of the serially diluted samples were
each inoculated onto different sterile
MacConkey agar plates in triplicates, the
inoculums were then spread evenly on the
surface of the media using a sterile spreader.
This was followed by incubation at 37oC for
24 hours, after which the colonies were
counted and the mean total coliform count
expressed as cfu/ml and cfu/g as applicable.
Total
Salmonella
-
Shige
lla
counts
This was determined with the
Salmonella
-
Shigella
agar using the spread plate method
as described by Prescott et al. (2005). One
milliliter of the serially diluted samples was
inoculated onto sterile pre-dried Salmonella-
Shigella
agar plates in duplicates. The
inocula were then spread evenly on the
surface of the media using a sterile spreader.
The plates were then incubated at 37oC for
24 hours, after which the colonies that
developed were counted and the mean total
Salmonella
-
Shigella
counts re
corded
accordingly for water, sediment, soil and
waste water samples.
Total
Vibrio
count
Total
Vibrio
count was determined with the
thiosulphate citrate bile salt (TCBS) agar
using the spread plate technique as described
by Prescott et al (2005). One milliliter of the
serially diluted samples were inoculated
onto sterile pre-dried TCBS agar plates in
triplicates and then spread evenly with a
sterile bent glass rod. The plates were
incubated at 37oC for 24 hours, after which
the colonies that developed were counted
and the mean recorded accordingly for
surface water, wastewater, sediment and soil
samples.
Total fungal counts
This was determined using the potato
dextrose agar (PDA) onto which sterile
streptomycin (50 mg/ml) had been added to
suppress bacterial growth (Okerentugba and
Ezereonye, 2003). The spread plate
technique as described by Prescott et al
(2005) was adopted. An aliquot (0.1ml) of
the serially diluted samples were inoculated
in triplicates onto sterile pre-dried PDA
plates and then spread evenly with a sterile
glass spreader. The plates were incubated at
room temperature for about 3-5 days after
which the colonies were counted and the
mean of the count recorded accordingly.
Total hydrocarbon utilizing bacterial
counts.
The population of hydrocarbon utilizing
bacteria was determined by inoculating
0.1ml aliquot of the serially diluted samples
onto mineral salt agar media using the
spread plate technique as described by
Odokuma (2003). The vapour phase transfer
method of Mills and Colwell (1978) was
adopted. It employed the use of sterile filter
paper discs soaked in filter-sterilized crude
oil which served as the only carbon source
in the mineral salt agar. The sterile crude
oil
-soaked filter papers were then aseptically
transferred to the inside covers of the
inoculated petri dishes and incubated for 5
days at room temperature. After the
incubation period, mean of the colonies
were recorded.
1188
Fig.
1
Map of Imo and Rivers states showing the study areas
1189
Table
.1
Identification of Samp
ling stations, points, coordinates and sample types in the study areas
Sampling point Co
-ordinates
Sampling
Stations
Sampling
Points
Northing (N)
Easting (E)
Types of Samples
A
05
0
28.432
/
007
0
03.200
/
Soil (Test sample)
B
05
0
28.441
/
007
0
03.209
/
Soil (Test sample)
C
05
0
28.582
/
007
0
03.312
/
Surface water and Sediment
(Test samples)
Egbu Abattoir I
D
05
0
28.559
/
007
0
3.231
/
Soil (Control sample)
A 050
29.651
/
007
0
04.205
/
Waste water
B
05
0
29.668
/
007
0
04.215
/
Waste water
C
05
0
29.705
/
007
0
04.285
/
Waste water
Egbu Abattoir II
Otamiri River
D
A
B
05
0
29.802
05
0/
28.426
05
027.423
007
0
04.918
/
007
003.179
007
004.156
Waste water
Surface water and
Sediment(Test sample)
Surface water and Sediment
(control)
A
04
0
48.886
/
007
0
2.707
/
Soil (Test sample)
B
04
0
48.782
/
007
0
2.608
/
Soil (Test sample)
C
04
0
48.615
/
007
0
2.405
/
Surface water and Sediment
(Test samples)
Trans
-
Amadi Abattoir I
D
04
0
48.442
/
007
0
2.303
/
Soil (Control sample)
A
04
0
49.789
/
007
0
03.801
/
Waste water
B
04
0
49.628
/
007
0
03.702
/
Waste water
C
04
0
49.522
/
007
0
03.665
/
Waste w
ater
Trans
-
Amadi abattoir II
Oginigba Creek
D
A
B
04
0
49.444
04
0/
50.001
04
050.111
007
0
03.545
/
007
004.425
007
0
04.225
Waste water
Surface water and Sediment
(Test samples)
Surface water and Sediment
(Control)
Total hydrocarbon utilizing fungal counts
Total hydrocarbon utilizing fungi was
determined by inoculating 0.1ml of the
serially diluted samples onto mineral salt
agar using the method of Odokuma (2003).
Eight hundred milliliter of the mineral salt
medium was supplemented with 70mg of
Aureomycin hydrochloride in 200ml of
steri
le distilled water (Odokuma, 2003). The
vapour phase transfer method of Mills and
Colwell (1978) was adopted. It employed
the use of sterile filter paper discs soaked in
filter
-sterilized crude oil which served as the
only carbon source in the mineral salt agar.
The sterile crude oil-soaked filter papers
were then aseptically transferred to the
inside covers of the inoculated petri dishes
and incubated for 5-10 days at room
temperature. After the incubation period,
mean of the colonies for the triplicate plates
were calculated and recorded accordingly.
1190
Results and Discussion
The results of the microbial counts obtained
during the study period for Egbu abattoir in
the rainy season are presented in Table 1 for
the test and control samples respectively.
Tot
al heterotrophic count of 2.5 x 10
6
cfu/g
was obtained from the sediment sample
collected from Otamiri River in Egbu while
the waste water sample had the least
observed count of 1.2 x 106 cfu/ml during
this period. Results further showed that the
total fungal count of 6.0 x 10
5
cfu/g was
obtained for the sediment sample which was
observed to be significantly different from
those obtained from soil, waste water and
surface water samples (Table 1). Statistical
analysis using ANOVA revealed a
significant difference at 0.05 level of
significance in the total heterotrophic
bacterial count from sediment and those
from soil, waste water and surface water
samples.
Total hydrocarbon utilizing bacterial counts
gave the highest and least values of 7.4 x 105
cfu/ml and 3.0 x 10
5
cfu/ml
from surface
water and waste water, respectively. While
there was no significant difference in the
total hydrocarbon utilizing bacterial count
values obtained for sediment and waste
water samples, there were significant
differences between total hydrocarbon
utilizing bacterial count from surface water
and that from waste water, sediment and soil
at 0.05 level of significance. Total
hydrocarbon utilizing fungal count recorded
the highest and least values of 2.5x10
5
cfu/ml and 1.8x10
5
cfu/g, respectively from
surface water and soil samples. There was
no significant difference observed in the
values obtained from soil, sediment, waste
water and surface water. There were
significant differences in the total coliform
counts from the soil and surface water
samples analyzed accounting for the highest
and lowest values of 2.0x10
6
cfu/g and
1.210
6
cfu/ml, respectively. Salmonella
and
Shigella
counts revealed that there was no
significant difference between values of
1.0x10
6
cfu/g and 9.0x10
5
cfu/ml o
btained
for soil and waste water, respectively.
Likewise there was no significant difference
between the counts from sediment and
surface water samples. Analysis also showed
that while there was no significant
difference in the total
Vibrio
counts of
3.0x10
5
cfu/g and 3.5x10
5
cfu/g obtained
from soil and sediment, respectively,
significant difference existed between these
counts and that obtained from surface water.
From the test results in the dry season, soil
sample was found to be richer in total
hetero
trophic bacteria while surface water
had the least (Table 2). Results of analysis
of variance (ANOVA) indicated a
significant difference in the total
heterotrophic bacterial count at 0.05level of
significance between the samples analyzed.
There was no significant difference in the
total fungal counts of 3.0x105 cfu/g and
3.2x105 cfu/g from soil and sediment
samples, respectively. Surface water had the
highest fungal count of 6.0x10
5
cfu/ml,
which was found to be significantly different
from the values obtained from other
samples. Total hydrocarbon utilizing
bacterial count was found to have the
highest value of 1.0x10
6
cfu/g in the
sediment sample which was revealed
through statistical analysis (ANOVA) at
0.05 levels to be significantly different from
the values obtained for soil, surface water
and waste water samples. Total hydrocarbon
utilizing fungal count was observed to be
more in the surface water with a value of
2.5x10
5
cfu/ml, while waste water had the
least value of 8.0x10
4
cfu/ml. Analysis of
varian
ce (ANOVA) also showed a
significant difference in the values obtained.
1191
Table
.1
Microbial counts of samples contaminated by Egbu abattoir wastes for Rainy season
THBC
TFC
THUB
THUF
TCC
SSC
TVC
SAMPLES
Test
Control
Test
Control
Test
Control
Test
Con
trol
Test
Control
Test
Control
Test
Control
SOIL (cfu/g)
c
1.30X10
6
1.6X10
6
b
4.5X1O
5
4.0X10
5
b
5.4X10
5
3.0X10
5
a
1.8X10
5
2.5X10
5
a
2.0X10
6
8.0X10
5
a
1.0x10
6
3.0X10
5
a
3.0x10
5
2.5X10
5
SEDIMENT
(cfu/g)
a
2.5x10
6
2.0X10
6
a
6.0x10
5
5.1X10
5
c
3.5x10
5
3.0X10
5
a
2.0x10
5
3.0X10
5
b
1.5x10
6
1.0X10
6
b
4.0x10
5
3.0X10
5
a
3.5x10
5
3.0X10
5
WASTE
W
ATER
(cfu/ml)
c
1.2x10
6
9.5X10
5
c
2.5x10
5
3.0X10
5
c
3.0x10
5
2.5X10
5
a
2.0x10
5
2.5X10
5
d
1.0x10
6
6.0X10
5
a
9.0x10
5
0.00
c
0.00
0.00
SURFACE
WATER
(cfu/ml)
b
2.0x10
6
1.5X10
6
b
4.0x10
5
3.0X10
5
a
7.4x10
5
0.00
a
2.5x10
5
2.0X10
5
c
1.2x10
6
7.0X10
5
b
3.0x10
5
3.0X10
5
b
1.3x10
5
0.00
Means in the same column with the same letter are not significantly diff
erent at 5% level of significance according to LSD test.
Table
.2 Microbial counts of samples contaminated by Egbu abattoir wastes for Dry season
THBC
TFC
THUB
THUF
TCC
SSC
TVC
SAMPLES
Test
control
Test
control
Test
control
Test
control
Test
control
T
est
control
Test
control
SOIL (cfu/g)
a
2.8X10
6
1.9X10
6
b
3.0X10
5
3.0X10
5
c
3.6X10
5
3.1X10
5
b
1.8X10
5
1.0X10
5
b
1.5X10
6
1.1X10
6
a
8.0X10
5
4.0X10
5
b
2.5X10
5
3.0X10
5
SEDIMENT
(cfu/g)
b
2.5X10
6
2.0X10
6
b
3.2X10
5
4.5X10
5
a
1.0X10
6
6.5X10
5
c
1.0X10
5
1.5X10
5
c
1.0X10
6
7.6X10
5
b
2.5X10
5
3.0X10
5
a
3.5X10
5
2.5X10
5
WASTE
WATER
(cfu/ml)
c
2.2X10
6
1.7X10
6
c
1.5X10
5
0.00
d
2.0X10
5
3.5X10
5
d
8.0X10
4
0.00
a
2.0X10
6
1.0X10
6
c
1.2X10
5
2.5X10
5
c
1.0X10
5
0.00
SURFACE
WATER
(cfu/ml)
d
2.0X10
6
1.5X10
6
a
6.0X10
5
2.5X10
5
b
7.4X10
5
5.0X10
5
a
2.5X10
5
2.5X10
5
d
3.4X10
5
5.0X10
5
c
1.0X10
5
3.0X10
5
c
1.0X10
5
0.00
Means in the same column with the same lette
r are not significantly different at 5% level of significance according to LSD test.
1192
0
5
10
15
20
25
Acinetobacter
sp.
Bacillus sp.
Citrobacter sp.
Enterobacter
sp.
Escherichia sp.
Flavobacterium
sp.
Klebsiella sp.
Lactobacillus
sp.
Micrococcus
sp.
Proteus sp.
Pseudomonas
sp.
Salmonella sp.
Serratia sp.
Shigella sp.
Staphylococcus
sp.
Streptococcus
sp.
Vibrio sp.
Waste water
Soil
Sediment
Surface water
Percentage occurrence of bacterial isolates (%)
Fig
.1
Percentage occurrence of bacterial isolates from samples contaminated by Egbu abattoir wastes for Rainy Season
0
5
10
15
20
Bacillus sp.
Citrobacter sp.
Enterobacter
sp.
Escherichia sp.
Flavobacterium
sp.
Klebsiella sp.
Lactobacillus
sp.
Micrococcus
sp.
Proteus sp.
Pseudomonas
sp.
Salmonella sp.
Serratia sp.
Shigella sp.
Staphylococcus
sp.
Streptococcus
sp.
Vibrio sp.
Waste water Soil Sediment Surface water
Percentage occurrence of bacterial
isolates (%)
Bacterial
isolates
Fig.2
Percentage occurrence of bacterial isolates from samples contaminated by Egbu abattoir wastes for Dry Season
1193
Table
.3
Microbial counts of samples contaminated by Trans
-
Amadi abattoir wastes for rainy sea
son
THBC
TFC
THUB
THUF
TCC
SSC
TVC
SAMPLES
Test Control Test
control
Test
control
Test
control
Test
control
Test
control
Test
control
SOIL (cfu/g)
a
2.8X10
7
2.2X10
7
a
1.3X10
7
1.0X10
7
b
5.2X10
6
3.5X10
6
ba
1.1X10
6
2.0X10
6
a
2.5X10
7
1.4X10
7
b
9.5X10
6
4.0X10
6
a
6.0X10
6
3.0X10
6
SEDIMENT
(cfu/g)
a
3.0X10
7
2.5X10
7
b
8.0X10
6
5.0X10
6
d
1.3X10
5
6.0X10
6
a
2.
0X10
6
2.0X10
6
c
2.1X10
7
1.2X10
7
c
6.0X10
6
4.0X10
6
c
2.0X10
6
3.0X10
6
WASTE
WATER
(cfu/ml)
a
2.1X10
7
1.0X10
7
c
3.0X10
6
3.0X10
6
c
3.3X10
6
2.5X10
6
b
1.0X10
6
0.00
d
1.5X10
7
5.0X10
6
d
4.0X10
6
3.0X10
6
c
1.0X10
6
0.00
SURFACE
WATER
(cfu/ml)
a
2.8X10
7
2.4X10
7
c
3.0X10
6
4.0X10
6
a
1.0X10
7
6.0X10
6
b
9.0X10
5
1.5X10
6
b
2.5X10
7
8.0X10
6
a
1.5X10
7
7.0X10
6
b
3.6X10
6
3.2X10
6
Means in the same column with the same letter are not significantly different at 5% level of significance according to LSD tes
Table
.4
Microbial counts of sa
mples contaminated by Trans-
Amadi abattoir wastes for dry season
THBC
TFC
THUB
THUF
TCC
SSC
TVC
SAMPLES
Test
Control
Test
control
Test
control
Test
control
Test
control
Test
control
Test
control
SOIL (cfu/g)
a
3.0X10
7
2.5X10
7
a
1.5X10
7
1.1X10
7
d
3.2X10
6
3.0X10
6
b
3.5X10
6
3.0X10
6
a
2.9X10
7
1.2X10
7
a
1.7X10
7
8.0X10
6
a
5.0X10
6
3.5X10
6
SEDIMENT
(cfu/g)
a
2.8X10
7
2.2X10
7
b
1.2X10
7
1.0X10
7
a
1.4X10
7
6.0X10
6
a
5.0X10
6
3.5X10
6
a
2.5X10
7
1.0X10
7
b
9.4X10
6
6.0X10
6
a
5.0X10
6
3.0X10
6
WASTE
WATER
(
cfu/ml)
a
2.5X10
7
1.5X10
7
c
3.5X10
6
3.0X10
6
c
5.0X10
6
4.0X10
6
c
2.0X10
6
2.0X10
6
b
1.1X10
7
7.5X10
6
c
2.5X10
6
3.0X10
6
b
1.5X10
6
0.00
SURFA
CE
WATER
(cfu/ml)
a
2.5X10
7
2.0X10
7
c
3.0X10
6
3.0X10
6
b
8.5X10
6
7.0X10
6
d
1.8X10
5
2.0X10
6
ab
2.0X10
7
9.0X10
6
b
1.0X10
7
5.0X10
6
b
2.1X10
6 3
.0X10
6
Means in the same column with the same letter are not significantly different at 5% level of significance according to LSD test.
1194
While waste water had the highest value for
total coliform count of 2.0x10
6
cfu/ml,
surface water samples had the least count of
3.4x10
5
cfu/ml. It was also observed that
waste water and surface water samples
showed no significant difference in the
S
almonella
-
Shigella
and total
Vibrio
counts.
But while
Salmonella
-
Shigella
count had the
highest value of 8.0x10
5
cfu/g from the soil
samples, sediment sample had the highest
Vibrio
count of 3.5x10
5
cfu/g.
T-test revealed that there were significant
differences between the total heterotrophic
bacterial counts of test and control samples
from sediment and surface water samples,
soil and waste water samples. There was no
significant difference however in the total
fungal counts of tests and controls of
sediment and soil samples from Otamiri
River. While there was difference
statistically at 0.05 level of significance
using the t-test in the total hydrocarbon
utilizing bacterial counts obtained from the
test and control samples of sediment, soil
and surface water.
Results of Microbial counts obtained from
samples collected from Trans-
Amadi
abattoir during the rainy season are
pres
ented in Table 3. From the test results,
sediment samples from Oginigba Creek in
Trans Amadi had the highest values for total
heterotrophic bacterial count and total
hydrocarbon utilizing fungal count of
3.0x107 cfu/g and 2.0x106 cfu/g,
respectively. Further analysis revealed that
the soil and surface water had more and
equal levels of total heterotrophic bacterial
and total coliform counts than other
samples. While the soil had higher levels of
total fungal count and total V
ibrio
count of
1.3x10
7
cfu/g and 6.0x106 cfu /g,
respectively,
Salmonella
-
Shigella
count had
higher value of 1.5x10
7 cfu/ml in the surface
water samples. In all, waste water had the
least values for all the microbial groups
enumerated. It was further ascertained
through statistical analysis (ANOVA) at
0.05 confidence limit that while there was
no statistical difference in the total
heterotrophic bacterial counts values from
the samples, there was a statistical
difference in the values obtained for total
hydrocarbon utilizing bacterial, to
tal
coliform and
Sallmonella
-
Shigella
counts.
However, while there was statistical
difference between total fungal and total
Vibrio
counts from soil and other samples,
there was no statistical difference in the total
fungal counts from the waste water and
surface water samples. The same was
applicable to the total
Vibrio
counts
obtained from sediment and waste water.
T-test at 0.05 confidence limit showed that
there was no statistical difference in the total
heterotrophic and total hydrocarbon utilizing
bacterial counts from test and control of
sediment samples from Oginigba Creek,
while there was a significant difference in
the counts from soil samples, surface water
and waste water samples. Total fungal
counts from sediment and soil samples
showed significant difference, while that
from surface water and waste water samples
did not show any significant difference.
From the Trans Amadi abattoir, the soil
samples had more total heterotrophic
bacteria, total fungi and total
Vibrio counts
of 3.0x10
7
cfu/g, 1.5x10
7
cfu/g and 5.0x106
cfu/g, respectively while total hydrocarbon
utilizing bacteria had 3.2x106 cfu/g in the
dry season. Lower counts of total
heterotrophic bacteria, total fungi, total
coliform, S
almonella
-
Shigella
and total
V
ibrio
counts, were recorded for both
sediment and surface water samples, while
waste water also had low values for all the
microbial groups enumerated within the
same abattoir. There was statistical
difference in the total coliform count value
from waste water and those from soil and
1195
sediment samples. Furthermore, total V
ibrio
counts from the soil and sediment showed
no significant difference; likewise no
significant difference existed between total
V
ibrio
count from the waste water and that
from surface water. Microbial count
results
also indicated statistical difference in the
total hydrocarbon utilizing bacterial values
obtained from test and control samples of
sediment, while counts from test and control
samples of soil, surface water and waste
water did not show any signifi
cant
difference.
Microorganisms are said to be ubiquitous
and are known for essential functions which
include; decomposition of organic materials,
bioaccumulation of chemicals and
biogeochemical cycling of elements. Their
presence, abundance and growth in the
environment are greatly influenced by
factors such as pH, temperature, pressure,
availability of nutrients and salinity
(Ogbonna and Ideriah, 2014).
The result in Table 1 revealed that sediment
from the Otamiri River (contaminated by
Egbu abattoir) contained more total
heterotrophic bacteria and fungi than the
soil, waste water and surface water during
the rainy season. This might be as a result of
increased microbial load washed into the
river from the soil by the rain and the fact
that more nutrien
ts are brought in by the rain
through leaching of the soil which
eventually settles at the bottom of the river,
leading to increased nutrient levels which
encouraged rapid multiplication of bacteria
and fungi present. During the dry season, the
soil had higher total heterotrophic bacterial
count of 2.8x10
6
cfu/g than the other
samples. This, according to Adesemoye
et
al
(2006) could be as a result of
destabilization of the soil ecological balance
arising from contamination with abattoir
wastes. Another pos
sible reason for this high
bacteria count in the soil during this period
could be as a result of accumulation of
wastes which are sources of microbial
nutrients, since there was no rain to wash
them off. The surface water had total
heterotrophic fungal count of 6.0x105 cfu/ml
which was higher than that from other
samples. This could probably be due to high
contamination of the river by air-
borne
fungal spores. Generally, the counts of
bacteria and fungi from sediment, waste
water and surface water samples from the
test samples are higher than those from
controls which signified contamination of
the test samples by untreated abattoir
wastes. However, counts of bacteria and
fungi from test and control soil samples
were almost equal which could be as a result
of droppings from cattle that move around
the abattoir and within the neighbourhood.
Coliforms were isolated from all the samples
collected from the two abattoirs. The
presence of this physiologic group in these
samples is an indication of feacal
contam
ination of the samples (Prescott
et
al.,
2005). This is possible since the cow
dung is indiscriminately deposited within
and around the abattoir. Through surface
run
-off, some of the feacal materials are
carried to the nearby water body, leading to
the presence of coliforms in such water
body. Statistically, difference existed at 0.05
confidence limit in the level of total
coliforms in all the samples from Egbu in
the rainy and dry seasons, than the Trans-
Amadi abattoir in the rainy season, this
could be as a result of uneven contamination
of these samples with fecal materials from
the cows during these periods. However, the
situation was not the same with the level of
coliform counts in the samples from Trans-
Amadi abattoir in the dry season. This could
be as a result of even contamination of the
samples with cow dung.
1196
0
5
10
15
20
25
Achromobacter
sp.
Acinetobacter
sp.
Bacillus sp.
Enterobacter
sp.
Escherichia sp.
Flavobacterium
sp.
Klebsiella sp.
Micrococcus
sp.
Proteus sp.
Pseudomonas
sp.
Salmonella sp.
Serratia sp.
Shigella sp.
Staphylococcus
sp.
Streptococcus
sp.
Vibrio sp.
Waste water
Soil
Sediment
Surface water
Percentage occurrence of bacterial isolates
(%)
Bacterial isolates
Fig
.3
Percentage occurrence of bacterial isolates from samples contaminated by Trans Amadi abattoir wastes for rainy season
Fig
.4
Percentage occurrence
of bacterial isolates from samples contaminated by Trans Amadi abattoir wastes for dry season
0
2
4
6
8
10
12
14
16
18
Achromobacter
sp.
Acinetobacter
sp.
Bacillus sp.
Enterobacter sp.
Escherichia sp.
Flavobacterium
sp.
Klebsiella sp.
Micrococcus
sp.
Proteus sp.
Pseudomonas
sp.
Salmonella sp.
Serratia sp.
Shigella sp.
Staphylococcus
sp.
Streptococcus
sp.
Vibrio sp.
Waste water
Soil
Sediment
Surface water
Bacterial isolates
Percentage occurrence of bacterial isolates (%)
1197
Salmonella
and
Shigella
were present in the
samples collected from the abattoirs. Their
presence was not astonishing since they co-
habit with coliforms in the intestinal tract of
warm blooded animals. Cow dung could be
a good source of coliforms around the
abattoirs. Statistically, there was little or no
difference at p < 0.05 in the level of
Salmonella
-
Shigella
counts from all the
samples collected from the Otamiri River
and Ogingba creek abattoirs in the rainy and
dry seasons. This could be as a result of
even distribution of these organisms during
these seasons. The soil had higher counts
of 3.0x104cfu/g and 3.7x104 cfu/g,
respectively within the periods. This could
be due to random and uncontrolled
deposition of wastes including cow dung
within and around the abattoir environment.
There was statistical difference at 0.05
confidence limit in the level of
Salmonella
-
Shigella
counts from samples collected both
in the rainy and dry seasons from Trans-
Amadi abattoir. Surface water from
Oginigba Creek and soil samples had higher
counts of 1.5x107 cfu/ml and 1.7x104 cfu/g
in both the rainy and dry seasons
respectively. This can be as a result of
increased surface run-off from the abattoir
into the river in the rainy season and
accumulation of organic wastes including
cow dung in the soil around the abattoir in
the dry season respectively. However, there
was much difference in the counts of
Salmonella
-
Shigella
obtained in the test and
control samples from Trans-Amadi abattoir,
with the test samples giving higher counts
than the control samples. This is probably
due to domestic activities that take place at
different points around these abattoirs which
can equally be a means of contaminating the
environments outside these abattoirs with
pathogenic organisms like Salmonella
and
Shigella
.
Vibrio
was isolated from most of the
samples. This indicates that the sampling
points were impacted by human activities.
However,
Vibrio
was not isolated from
waste water from Egbu abattoir during the
rainy season. This is an indication that these
samples were not impacted by human
activities. It is equally possible that the
samples were contaminated with organic
wastes that were devoid of
Vibrio
species
The relatively high incidence of
Klebsiella,
Enterobacter, Escherichia, Salmonella,
Shigella, Citrobacter, Serratia and
Proteus
around the test sampling points may be
connected with high rate of cattle defecation
near the sites. The introductions of wastes
from the abattoir and the surface run
-
off into
the sites and nearby rivers during the rains
are also contributory factors (Ezeronye and
Ubalua, 2005). The presence of these
isolates in this study gives credence to these
findings. The isolation of
E.coli
and other
coliforms is an indication of recent human
contamination of the sampling points, and is
of great public health concern (Ezeama and
Nwamkpa, 2002).
The presence of
Bacillus
sp supports the finding by Ezeronye
and Ubalua (2005). The organism is mostly
a soil inhabitant and its presence could be as
a result of contamination from overland run-
off. The presence of Pseudomonas,
Acinetobacter, and Lactobacillus around the
abattoir is possible since they have been
reported to be agents of meat spoilage
(Frazier and Westhoff, 2003). Occurrence of
Pseudomonas sp as a heterotrophic and
hydrocarbon utilizing bacteria has been
reported (Loureiro et al. 2005). The
presence of Pseudomonas sp. within the
abattoir environment is possible. This is
probably due to the presence of
hydrocarbons (PAHs) within the abattoir.
This observation supports the report by Faria
and Bharathi (2006) that Pseudomonas sp is
widespread in the environment and
concluded that they could contribute to the
1198
oxidation of hydrocarbons in the
environment. The same reason is applicable
to
Achromobacter
and Acinetobacter, which
according to Leahy and Colwell (1990) are
among the hydrocarbon degraders. The
incidence of
Staphylococcus
in this study is
in agreement with the report by Chen et al
(2001) who reported that
Staphylococcus
is
naturally found in the hides of cattles. The
recovery of Flavobacterium which is said to
be authochthonous to the environment,
agrees with the report of Austin (1988). The
isolation of
Streptococcus
sp. and
Micrococcus sp in this study agrees with the
report of Adeyemo et al (2002), who
isolated these organisms from abattoir
environments.
Abattoir wastes pollution has adverse
impacts on aquatic environment thus
triggering algal blooms (eutrophication),
depletion of dissolved oxygen, destruction
of habitat and fish kills, thereby reducing the
population of fishes and other aquatic
organisms (Meadows, 1995; Chukwu et al
.,
2008). Some of the consequences of abattoir
pollution are transmission of diseases by
water borne pathogens, eutrophication of
natural water bodies, accumulation of toxic
or recalcitrant chemicals in the soil,
destabilization of ecological balance and
negative effects on human health
(McLaughlin and Mineau, 1995; Sinha,
1997; Bridges et val., 2000; Boadi and
Kuitunen, 2003; Amisu et al., 2003).
Potential health risks from waterborne
pathogens can exist in water contaminated
by abattoir effluents (Cadmus et al., 1999),
runoff from feedlots (Miner et al., 1966),
dairy farms (Janzen et al., 1974), grazed
pastures (Doran and Linn 1979; Kunkel
et
al
., 1983), fallow and sod amended with
poultry litter (Giddens and Barnet, 1980),
grassland treated with dairy manure
(McCaskey
et al., 1971), and sewage sludge
treated land (Dunigan and Dick, 1980).
Po
llution of water resources by abattoir
wastes might lead to destruction of primary
producers and this in turn leads to
diminishing consumer populations in water.
The direct repercussion of this is
diminishing fish yield hence human diet
suffers. Therefore, continuous monitoring
should be maintained in order to promote
and maintain a safe working environment
and ensure detection when abnormalities
that could endanger both workers and
environment occur.
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